COMPETITION BETWEEN BACULOVIRUS POLYHEDRON AND P10 GENE-EXPRESSION DURING INFECTION OF INSECT CELLS
COMPETITION BETWEEN BACULOVIRUS POLYHEDRON AND P10 GENE-EXPRESSION DURING INFECTION OF INSECT CELLS
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DOI：
10.1128/jvi.67.5.2664-2671.1993
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发表时间：
1993-05-01
影响因子：
5.4
通讯作者：
CERUTTI, M
中科院分区：
文献类型：
--
作者：
CHAABIHI, H;OGLIASTRO, MH;CERUTTI, M
多角体蛋白和p10基因在感染后期同时表达。为了确定这两个基因之间是否在转录和/或翻译水平上发生任何竞争，构建了一系列缺失p10或多角体蛋白基因的启动子和编码序列的苜蓿银纹夜蛾核型多角体重组病毒。制备了两种仅具有一种极晚期启动子的修饰杆状病毒，即单晚期启动子病毒AcSLP 10和AcSLP 33。多角体蛋白基因被用作报告基因，以允许p10和多角体蛋白驱动的表达之间的直接比较。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析和[S-35]甲硫氨酸标记实验表明，多角体蛋白的合成增强杆状病毒感染的细胞携带只有一个非常晚的启动子。然而，当多角体蛋白基因位于其天然位点时，这种增强更显着。用总细胞质RNA进行的斑点杂交实验表明，p10启动子的缺失导致来自多角体蛋白启动子的mRNA的增加。相反，当多角体蛋白基因启动子被删除时，没有检测到p10促进的mRNA的增加。
Polyhedrin and p10 genes are expressed concurrently during the late stage of infection. To determine whether any competition occurs between these two genes at a transcriptional and/or translational level, a series of Autographa californica nuclear polyhedrosis recombinant viruses with deletions of promoter and coding sequences of the p10 or polyhedrin gene was constructed. Two modified baculoviruses with only one of the very late promoters, single late-promoter viruses AcSLP10 and AcSLP33, were made. The polyhedrin gene was used as a reporter gene to allow direct comparison between p10 and polyhedrin-driven expression. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and [S-35]methionine labeling experiments showed that polyhedrin synthesis is enhanced in cells infected with baculoviruses carrying only one very late promoter. However, this enhancement is more significant when the polyhedrin gene is located in its natural site. Dot blot hybridization experiments carried out with total cytoplasmic RNA showed that deletion of the p10 promoter resulted in an increase of mRNAs derived from the polyhedrin promoter. In contrast, no increase of p10-promoted mRNA was detected when the polyhedrin gene promoter was deleted.